CAJ | 학술논문

目的：克隆、表达发热伴血小板减少综合征布尼亚病毒（SFTSV）非结构蛋白 NSs，对其功能初步鉴定。方法采用 PCR 法扩增经密码子优化后的 SFTSV NSs 基因，并克隆至 PGEX-4T-2载体中，构建原核表达载体 PGEX-4T-2-NSs，经酶切、测序鉴定后转化表达菌 BL21（DE3），再经诱导、纯化得到谷胱甘肽 S-转移酶（GST）-NSs 融合蛋白，用 Western Blot 验证融合蛋白 GST-NSs 的抗原性。结果成功构建了 GST-NSs 融合蛋白原核表达载体，并在 BL21细菌中获得高效表达；纯化后的融合蛋白具有良好的抗原性。结论实现了 SFTSV 重组 NSs 蛋白的原核高效表达，为进一步深入研究其结构与功能奠定了基础。
목적：극륭、표체발열반혈소판감소종합정포니아병독（SFTSV）비결구단백 NSs，대기공능초보감정。방법채용 PCR 법확증경밀마자우화후적 SFTSV NSs 기인，병극륭지 PGEX-4T-2재체중，구건원핵표체재체 PGEX-4T-2-NSs，경매절、측서감정후전화표체균 BL21（DE3），재경유도、순화득도곡광감태 S-전이매（GST）-NSs 융합단백，용 Western Blot 험증융합단백 GST-NSs 적항원성。결과성공구건료 GST-NSs 융합단백원핵표체재체，병재 BL21세균중획득고효표체；순화후적융합단백구유량호적항원성。결론실현료 SFTSV 중조 NSs 단백적원핵고효표체，위진일보심입연구기결구여공능전정료기출。
Objective To clone and express non-structural protein(NSs)of severe fever with thrombocytopenia syndrome bunyavirus (SFTSV);to characterize preliminary function of recombinant protein.Methods SFTSV NSs coding region was optimized and amplified by PCR,which was cloned into PGEX-4T-2 vector and validated by restriction digestion and sequence analysis.Confirmed expression vector was transformed into E.coli host strain BL21(DE3),which was cultured and induced for expression of glutathione S-transferase(GST)-NSs recombinant protein .GST-NSs recombinant protein was purified and tested for its antigenicity by Western Blot analysis.Results Prokayotic GST-NSs expression vector was constructed successfully. GST-NSs recombinant protein was expressed very effectively in BL21 bacteria cultures.Purified recombinant GST-NSs main-tained good antigenicity.Conclusion SFTSV NSs recombinant protein was expressed in prokaryotic system with high efficien-cy ,which laid a solid foundation for further study of its structure and functions .